METHODS FOR BIOLOGICAL SAMPLE PROCESSING AND ANALYSIS
Provided are methods for biological sample processing and analysis. A method can comprise providing a substrate configured to rotate. The substrate can comprise an array having immobilized thereto a biological analyte. A solution comprising a plurality of probes may be directed, via centrifugal force, across the substrate during rotation of the substrate, to couple at least one of the plurality of probes with the biological analyte. A detector can be configured to detect a signal from the at least one probe coupled to the biological analyte, thereby analyzing the biological analyte.
1 . A method for sequencing a nucleic acid molecule, said method comprising:
(a) providing an array of nucleic acid molecules adjacent to an open surface;
(b) dispersing a solution over said open surface to provide a layer of said solution over said open surface, wherein said solution comprises reagents including at least one nucleotide present under conditions sufficient to incorporate said nucleotide into a growing nucleic acid strand that is complementary to a nucleic acid molecule of said array of nucleic acid molecules; and
(c) detecting one or more signals or signal changes from said nucleic acid molecule, which one or more signals or signal changes are indicative of said nucleotide having incorporated into said growing nucleic acid strand.
2 . The method of claim 1 , wherein said open surface is accessible at any point in said open surface from a direction normal to a plane of said open surface.
3 . The method of claim 1 , wherein said open surface is not a flow cell.
4 . The method of claim 1 , wherein said open surface is substantially planar.
5 . The method of claim 1 , wherein said open surface is textured or patterned.
6 . The method of claim 1 , wherein said layer has a thickness of less than about 100 micrometers (μm) over said open surface.
7 . The method of claim 1 , wherein said solution comprises a plurality of nucleotides of a first canonical base type, including said at least one nucleotide.
8 . The method of claim 7 , wherein said solution comprises a first plurality of nucleotides of a first canonical base type, including said at least one nucleotide, and a second plurality of nucleotides of a second canonical base type different than said first canonical base type.
9 . The method of claim 1 , further comprising, subsequent to (c), (i) dispersing an additional solution over said open surface to provide an additional layer of said solution over said open surface, wherein said additional solution comprises additional reagents including at least one additional nucleotide present under conditions sufficient to incorporate into said growing nucleic acid strand that is complementary to said nucleic acid molecule, and (ii) detecting one or more additional signals or signal changes from said nucleic acid molecule, which one or more additional signals or signal changes are indicative of said at least one additional nucleotide having incorporated into said growing nucleic acid strand.
10 . The method of claim 9 , wherein said at least one nucleotide is of a first canonical base type and wherein said at least one additional nucleotide is of a second canonical base type is different than said first canonical base type.
11 . The method of claim 9 , wherein said solution and said additional solution are dispensed to said open surface using different nozzles.
12 . The method of claim 1 , further comprising, subsequent to (b), dispersing an additional solution over said open surface to provide an additional layer of said solution over said open surface, wherein said additional solution comprises a washing reagent.
13 . The method of claim 1 , wherein said detecting in (c) is performed during rotation of said open surface.
14 . The method of claim 1 , wherein said dispersing in (b) comprises subjecting said open surface to motion.
15 . The method of claim 14 , wherein said motion comprises rotational motion.
16 . The method of claim 1 , wherein said dispersing in (b) comprises spraying said solution over said open surface.
17 . The method of claim 1 , wherein said open surface is contained in a chamber that insulates an internal environment from an external environment.
18 . The method of claim 17 , wherein said internal environment has a higher humidity than an ambient atmosphere.
19 . The method of claim 17 , wherein at least a portion of said internal environment is maintained at temperatures above a dewpoint to prevent condensation.
20 . The method of claim 17 , wherein at least a portion of said internal environment is maintained at temperatures below a dewpoint to collect condensation.
21 . The method of claim 1 , wherein said solution comprises a plurality of labeled nucleotides, including said at least one nucleotide.
22 . The method of claim 21 , further comprising, subsequent to (c), cleaving or modifying a label from said at least one nucleotide.
23 . The method of claim 1 , wherein said solution comprises a plurality of unlabeled nucleotides.
24 . The method of claim 1 , wherein said solution comprises a plurality of labeled nucleotides and a plurality of unlabeled nucleotides.
25 . The method of claim 1 , wherein said at least one nucleotide is not reversibly terminated.
26 . The method of claim 1 , wherein said at least one nucleotide is reversibly terminated.
27 . The method of claim 1 , wherein said one or more signals or signal changes comprise fluorescent signals or signal changes.
28 . The method of claim 1 , wherein said open surface comprises an array of individually addressable locations, and wherein said nucleic acid molecule is immobilized to an individually addressable location of said array of individually addressable locations.
29 . The method of claim 1 , wherein said nucleic acid molecule is coupled to a bead, which bead is immobilized to said open surface.
30 . The method of claim 29 , wherein said bead comprises a plurality of nucleic acid molecules, including said nucleic acid molecule, attached thereto, and wherein said plurality of nucleic acid molecules have sequence homology to one another.